Inhibition of Sporulation of Genus Bacillus by Various Microbial Protease Inhibitors

Several analogs modifying the 6-methoxy-2-methoxymethyl-3-(3,4-methyienedioxyphenyl)-1,4-benzodioxan-7-yl group of haedoxans were synthesized and their insecticidal activity was examined. 2-(2,6-Dimethoxyphenoxy)-1-hydroxy-6-(2-methoxy-5-methoxyethoxyphenyl)-3,7-dioxabicyclo-[3.3.0]octane, which lacked the 3-(3,4-methylenedioxybenzyloxy) moiety of the benzodioxanyl group, was not insecticidal, but caused prolonged paralysis of the housefly. A compound replacing the 6-(2-methoxy-5-methoxyethoxyphenyl) by 6-(5-butoxy-2-methoxyphenyl) exhibited insecticidal activity comparable to one thirty-thousandth of that of haedosan A. It became evident that the 1,4-benzodioxane framework charging the 3-(3,4-methylenedioxy)phenyl group is important for the insecticidal activity of haedoxans.     

Cell-Bound Serine Proteinase of Sporulating Cells of Bacillus subtilis

2-Deuterio-2-cyclohexen-l-one, 3-deuterio-2-cyclohexen-l-one and 2-methyl-2-cyclohexen-1-one were reduced by Clostridium La 1 giving a single bioconversion product resulting from reduction of the carbon-carbon double bond. Stereochemistry of the reaction was studied.     

Purification and Properties of an S-PI(Pepstatin Ac)-insensitive Carboxyl Proteinase from a Xanthomonas sp. Bacterium

A S-PI(Pepstatin Ac)-insensitive carboxyl proteinase was found in culture filtrate of a Xanthomonas sp. bacterium. The carboxyl proteinase was highly purified and about 100 mg of the enzyme was obtained from 601 of culture filtrate, with a recovery of 25%. The optimum condition for the action of the purified enzyme toward casein was approx. pH 2.7 and its activity was not inhibited by any of such carboxyl proteinase inhibitors as Pepstatin, S-PI, and DAN but EPNP inhibited it. Such behavior of the enzyme against inhibitors resembles that of Pseudomonas sp. carboxyl proteinase, the first found from a bacterium. Some differences were observed, however, in their properties such as optimum pH, isoelectric point, and amino acid composition.     

Streptomyces Pepsin Inhibitor-Insensitive Carboxyl Proteinase from Lentinus edodes

A Streptomyces-pepsin inhibitor (S-PI or Pepstatin Ac), and DAN-insensitive carboxyl proteinase was found in a still culture filtrate of Lentinus edodes. The new carboxyl proteinase was purified, and about 9 mg purified enzyme was obtained from 19 liters of culture filtrate, with 11% recovery. The enzyme showed a single band on polyacrylamide gel electrophoresis. The molecular weight and isoelectric point were 40,000 and pH 4.2, respectively. The enzyme did not contain histidine and was composed of 387 amino acid residues. The enzyme was most active between pH 2.7 ~ 2.9, and stable over a pH of 3.2 ~ 5.2 for 3 hr at 37°C. The enzyme was not inhibited by S-PI or synthetic pepsin inhibitors such as DAN and EPNP. The physicochemical and enzymological properties were very similar to those of Scytalidium lignicolum carboxyl proteinase A, which was reported to be an S-PI-, and DAN-insensitive carboxyl proteinase.     

Structures of the Laccase-catalyzed Oxidation Products of Hydroxy-benzoic Acids in the Presence of ABTS [2,2′-Azino-di-(3-ethylbenzothiazoline-6-sulfonic Acid)]

Esters selected from the combination of tricycloalkanecarboxylic acids and some representative pyrethroidal alcohols were prepared, and their insecticidal activities were measured by topical application on housefly, mosquito and German cockroach. Both the exo- and endo-isomers of 2,3-trimethylenenorbornane-2-carboxylic acid exhibited a high level of activity when esterified with m-phenoxymandelonitrile. Piperonyl butoxide showed a strong synergistic effect on some of the esters. Several structural resemblances were found among these active tricyclic acids and the chrysanthemummonocarboxylic acids, which were suggested to be related to insecticidal activity.     

A Facile Synthesis of 4,8-Dimethyldecanal,Aggregation Pheromone of Flour Beetles,and Its Analogues

A Rhodococcus erythropolis expression system for the bovine lactoferrin C-lobe was constructed. The DNA fragments encoding the BLF C-lobe were amplified and cloned into vector pTip LCH1.2. R. erythropolis carrying the pTip-C-lobe was cultured at 30 °C with shaking, and expression of the rBLF C-lobe was induced by adding 1 μg/ml (final concentration) thiostrepton. The rBLF C-lobe was isolated in native and denatured (8 M urea) form by Ni-NTA affinity chromatography. To obtain a bioactive rBLF C-lobe, the protein isolated in the denatured form was refolded by stepwise dialysis against refolding buffers. The antibacterial activity of the rBLF C-lobe was tested by the filter-disc plate assay method. The refolded rBLF C-lobe demonstrated antibacterial activity against selected strains of Escherichia coli.     

Anthranilic Acid,as a Fruiting Body Inducing Substance in Favolus arcularius,from a Strain TA 7 of Actinomycetes

The stereochemical inversion of (R)-5-hydroxymethyl-3-tert-butyl-2-oxazolidinone (la) or (R)-5-hydroxymethyl-3-isopropyl-2-oxazolidinone (lb) to the corresponding (S)-isomer was accomplished via a key intermediate, (R)-3-N-ethoxycarbonyl-N-tert-butylamino-l,2-epoxypropane (5a) or (R)-3-N-ethoxycarbonyl-N-isopropylamino-l,2-epoxypropane (5b), in a high enantiomeric excess. (S)-la (99%e.e.) or (S)-lb (91%e.e.) was thus obtained from the respective (R)-isomer (la; 99%e.e., lb; 95%e.e.).     

General Properties of Endo-N-acetylmuramidase of Bacillus subtilis YT-25

The enzymatic behaviour, amino acid composition and some physical properties of a new endo-N-acetylmuramidase (B-enzyme) of Bacillus subtilis YT–25 were determined and compared with hen’s egg white lysozyme. The molecular weight was estimated to be about 13000 by the sedimentation equilibrium method. The isoelectric point was pH 9.8. The amino acid composition indicates that the enzyme is rich in basic amino acids, especially lysin. Maximal activity on the lysis of cell walls of M. lysodeikticus occurred at pH 6.2. The enzyme was stable at pH 3.5 ~ 6.0. The specific activity for the lysis of cell walls of M. lysodeikticus was less than fourth part of that of hen’s egg white lysozyme. Digest of cell walls of M. lysodeikticus with B-enzyme consisted greater numbers of high molecular products than digest with egg white lysozyme. Substrate specificity of B-enzyme seemed to be different from that of egg white lysozyme.